WO2016149492A1 - Methods of banking placental tissue and cells - Google Patents

Methods of banking placental tissue and cells Download PDF

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Publication number
WO2016149492A1
WO2016149492A1 PCT/US2016/022865 US2016022865W WO2016149492A1 WO 2016149492 A1 WO2016149492 A1 WO 2016149492A1 US 2016022865 W US2016022865 W US 2016022865W WO 2016149492 A1 WO2016149492 A1 WO 2016149492A1
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WIPO (PCT)
Prior art keywords
placental
tissue
umbilical cord
cells
placenta
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PCT/US2016/022865
Other languages
French (fr)
Inventor
Ralph FARIELLO
Manoochehr KHORSHIDI
Jessica O'DONNELL
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Anthrogenesis Corporation
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Publication of WO2016149492A1 publication Critical patent/WO2016149492A1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0603—Embryonic cells ; Embryoid bodies
    • C12N5/0605—Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02—Preservation of living parts
    • A01N1/0205—Chemical aspects
    • A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
    • A01N1/0221—Freeze-process protecting agents, i.e. substances protecting cells from effects of the physical process, e.g. cryoprotectants, osmolarity regulators like oncotic agents
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02—Preservation of living parts
    • A01N1/0278—Physical preservation processes
    • A01N1/0289—Pressure processes, i.e. using a designated change in pressure over time
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/48—Reproductive organs
    • A61K35/50—Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells

Definitions

  • the disclosure herein relates to methods of banking placental tissue.
  • said placental perfusate has been collected from a placenta that has been exsanguinated but not perfused to remove residual blood. In another embodiment, said placental perfusate has been collected from a placenta that has not been exsanguinated.
  • the placental tissue that is banked in accordance with the methods described herein is an exsanguinated placenta. In one embodiment of the methods described herein, the exsanguinated placenta is additionally banked with blood obtained from the exsanguinated placenta, e.g., whole cord blood.
  • the placental tissue that is banked in accordance with the methods described herein is an exsanguinated and perfused placenta.
  • an exsanguinated and perfused placenta is banked using the methods described herein, and blood obtained from the exsanguinated placenta, e.g., whole cord blood, and/or placental perfusate obtained from the placenta is/are banked with the exsanguinated and perfused placenta.
  • Methods of isolating blood/cells from an exsanguinated and/or perfused placenta are known in the art and described in Section 4.2.
  • said umbilical cord is banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), umbilical cord membrane, placental vessels, platelet-rich plasma from placental blood or umbilical cord blood, and isolated cells from the placenta (see Section 4.3).
  • placental perfusate cells isolated from placental perfusate, amniotic membrane, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), umbilical cord membrane, placental vessels, platelet-rich plasma from placental blood or umbilical cord blood, and isolated cells from the placenta (see Section 4.3).
  • the placental tissue that is banked in accordance with the methods described herein is umbilical cord membrane.
  • said umbilical cord membrane is decellularized.
  • said umbilical cord membrane is banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, chorionic membrane, umbilical cord, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), placental vessels, platelet-rich plasma from placental blood or umbilical cord blood, and/or isolated cells from the placenta (see Section 4.3).
  • the placental tissue that is banked in accordance with the methods described herein is one or more placental vessels.
  • said placental vessels are banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, chorionic membrane, umbilical cord, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), umbilical cord membrane, platelet-rich plasma from placental blood or umbilical cord blood, and/or isolated cells from the placenta (see Section 4.3).
  • the methods described herein comprise banking of (i) at least one unit of placental blood or umbilical cord blood and (ii) at least one unit of placental perfusate or total nucleated cells collected from placental perfusate, wherein said placental or umbilical cord blood and said placental perfusate or total nucleated cells collected from said placental perfusate are isolated from the same placenta.
  • the placental tissue banked in accordance with the methods described herein is allogeneic to the individual that has elected to bank the placental tissue, e.g., the customer of the placental tissue bank.
  • the placental tissue banked in accordance with the methods described herein is autologous to the individual that has elected to bank the placental tissue, e.g., the customer of the placental tissue bank.
  • the methods described herein comprise assigning the placental tissue to be banked an identifier based on the source of the placenta.
  • the methods described herein comprise assigning the placenta that is the source of placental tissue to be banked a first identifier, and assigning each placental tissue to be banked from the placenta a second tissue-specific identifier.
  • the second tissue-specific identifier incorporates said first identifier (for example, the placenta can be assigned the identifier XYZ, and amnion from the placenta can be assigned the identifier XYZ 123).
  • the placental tissue bank in which the placental tissue is banked comprises a system for storing and retrieving the identifiers, e.g., the first and second identifiers.
  • a method of providing placental tissue to a customer of the placental tissue bank comprises: (a) providing the customer a selection of placental tissue from said placenta or portion thereof to bank, (b) receiving an election from the customer as to which placental tissue to bank; (c) processing the placenta to isolate the elected tissue(s); and (d) banking the tissues at the direction of said customer, thereby providing placental tissue to a customer.
  • placental tissue derived from one or more sources (e.g., one or more placentas).
  • said placental tissue is autologous to a customer of the placental tissue bank that banks the placental tissue.
  • said placental tissue is heterologous to a customer of the placental tissue bank that banks the placental tissue.
  • the source of the placental tissue may be any placental animal (e.g., a primate, for example a human).
  • said placental tissue is derived from a human.
  • the methods described herein comprise the banking of placental tissue, and may additionally comprise the assignment of identifiers to the placental tissue to allow for subsequent monitoring and retrieval of the placental tissue after banking.
  • the placental tissue is assigned an identifier based on the region of the placenta from which said placental tissue is derived.
  • the placenta that is the source of placental tissue to be banked is assigned a first identifier, and each placental tissue to be obtained from the placenta (e.g., amnion, chorion, etc.) is assigned a second, tissue-specific, identifier specific for the particular tissue.
  • said second identifier incorporates said first identifier into a single identifier.
  • the placental tissue bank that banks the placental tissue comprises a system for storing and subsequently retrieving said identifiers, e.g., said first and second identifiers.
  • placental tissue is banked in a placental tissue bank, wherein the method comprises the steps of (a) receiving a placenta or portion thereof, (b) processing the placenta to isolate the placental tissue as described herein, and (c) subsequently banking the placental tissue, wherein prior to step (a) or step (b), a customer elects which placental tissue from said placenta to bank.
  • placental tissue is banked in a placental tissue bank according to a method comprising (a) receiving an election from a customer of the placental tissue bank as to which placental tissue(s) to bank; (b) receiving a placenta or portion thereof; (c) processing the placenta to isolate the placental tissue as described herein; and (d) subsequently banking the elected placental tissue.
  • the one or more placental tissue elected by a customer are assigned a third identifier to indicate election.
  • the one or more placental tissue not elected by the customer are assigned a fourth identifier to indicate non-election.
  • a customer may elect to bank an intact placenta or one or more of several types of placental tissue.
  • a method of providing placental tissue to a customer comprising providing the customer a selection of placental tissue from said placenta or portion thereof to a bank, wherein the tissues comprise one or both of whole placental perfusate and/or cells isolated from placental perfusate.
  • said tissues provided to a bank may comprise (1) one or both of whole placenta perfusate and/or cells isolated from placental perfusate and (2) additionally one or more of amniotic membrane, chorionic
  • the methods described herein comprise providing placental tissue to a customer comprising providing the customer a selection of one or more placental tissue as described herein, receiving an election from the customer as to which placental tissue to bank, processing the placenta to isolate the elected tissues as described herein, and banking the tissues at the direction of said customer.
  • the methods described herein may comprise additionally providing umbilical cord blood and/or placental blood to a customer.
  • said umbilical cord blood and/or placental blood are autologous to the customer.
  • said umbilical cord blood and/or placental blood are heterologous to the customer.
  • the customer selects the tissue for banking to be one or more of whole placenta, amniotic membrane, chorionic membrane, whole umbilical cord blood, umbilical cord vessels, umbilical cord membrane, placental vessels, and additionally comprising providing the customer with the option of decellularizing said placental tissue.
  • an intact placenta may be processed prior to its banking.
  • the processed placental tissue may be an intact placenta, e.g., an intact placenta that has been exsanguinated or an intact placenta that has been
  • the processed placental tissue may be placental perfusate ⁇ e.g., the placental perfusate isolated as described in Section 4.2.1).
  • the placental tissue consists of the total nucleated cells isolated from placental perfusate ⁇ e.g., total nucleated cells isolated from a placenta perfused according to the methods described in Section 4.2.1).
  • a method of banking placental tissue comprises receiving a placenta or a portion thereof, processing said placenta or portion thereof to isolate the placental tissue, and banking said placental tissue comprising placental perfusate or total nucleated cells isolated from said placental perfusate.
  • placental perfusate from a single placental perfusion comprises about 100 million to about 500 million nucleated cells.
  • a human placenta is recovered shortly after its expulsion after birth.
  • the placenta is recovered from a patient after informed consent and after a complete medical history of the patient is taken and is associated with the placenta.
  • the placenta is recovered from a patient after informed consent and after a complete medical history of the patient is taken and is associated with the placenta.
  • the tracking of medical history continues after delivery.
  • the intact placenta is cleaned prior to further processing and handling.
  • the intact placenta is handled to separate its constituent tissues and cell types after cleaning.
  • placental tissues are dissected and isolated ⁇ e.g., the amniotic membrane, the chorionic membrane, umbilical cord vessels, umbilical cord membrane, and placental vessels are dissected and isolated) after cleaning the intact placenta.
  • cells associated with the placenta are isolated ⁇ e.g., umbilical cord blood is isolated) after cleaning the intact placenta.
  • the intact placenta is not dissected after cleaning.
  • the intact placenta or placental tissues derived therefrom are cryopreserved according to the methods described herein (see Section 5.1.2).
  • placental perfusate is collected before banking placental tissue and/or cells derived therefrom.
  • the umbilical cord blood and placental blood are removed prior to recovery of perfusate.
  • the cord blood in the placenta is recovered.
  • the placenta prior to recovery of placental perfusate, is drained of cord blood and further processed, e.g., perfused, to remove residual blood.
  • the placenta prior to recovery of placental perfusate, is drained of cord blood, but is not further processed to remove residual blood.
  • the placenta prior to recovery of placental perfusate, the placenta is neither drained of cord blood nor processed to remove residual blood.
  • the placenta can be subjected to a conventional cord blood recovery process.
  • a needle or cannula is used, with the aid of gravity, to exsanguinate the placenta (see, e.g., Anderson, U.S. Patent No. 5,372,581; Hessel et al., U.S. Patent No. 5,415,665).
  • the needle or cannula is usually placed in the umbilical vein and the placenta can be gently massaged to aid in draining cord blood from the placenta.
  • Such cord blood recovery may be performed
  • the placenta is gravity drained without further manipulation so as to minimize tissue disruption during cord blood recovery.
  • a placenta is transported from the delivery or birthing room to another location, e.g., a laboratory, for recovery of cord blood and collection of perfusate.
  • the placenta can be transported in a sterile, thermally insulated transport device (maintaining the temperature of the placenta between 20-28°C), for example, by placing the placenta, in a sterile zip-lock plastic bag, which is then placed in an insulated container.
  • the placenta is transported in a cord blood collection kit substantially as described in U.S. Patent No.
  • the placenta prior to collection of the perfusate, can be stored under sterile conditions and at either room temperature or at a temperature of 5 to 25°C.
  • the placenta may be stored for a period of longer than forty eight hours, or for a period of four to twenty -four hours prior to perfusing the placenta to remove any residual cord blood.
  • the placenta can be stored in an anticoagulant solution at a temperature of 5°C to 25°C.
  • Suitable anticoagulant solutions are well known in the art.
  • a solution of heparin or warfarin sodium can be used.
  • the anticoagulant solution comprises a solution of heparin (e.g., 1% w/v in 1 : 1000 solution).
  • the exsanguinated placenta is stored for no more than 36 hours before placental perfusate is collected.
  • the placenta can be oriented in such a manner that the umbilical artery and umbilical vein are located at the highest point of the placenta.
  • the placenta can be perfused by passage of a perfusion solution through the placental vasculature, or through the placental vasculature and surrounding tissue.
  • the umbilical artery and the umbilical vein are connected simultaneously to a pipette that is connected via a flexible connector to a reservoir of the perfusion solution.
  • the perfusion solution is passed into the umbilical vein and artery.
  • Plastic tubing connected to a sterile collection reservoir, e.g., a blood bag such as a 250 mL collection bag, is then inserted into the placental vein.
  • a sterile collection reservoir e.g., a blood bag such as a 250 mL collection bag
  • the tubing connected to the pump is inserted into the placental vein, and tubes to a collection reservoir(s) are inserted into one or both of the placental arteries.
  • the placenta is then perfused with a volume of perfusion solution, e.g., about 750 ml of perfusion solution. Cells in the perfusate are then collected, e.g., by
  • the placenta can be perfused a plurality of times over the course of several hours or several days. Where the placenta is to be perfused a plurality of times, it may be maintained or cultured under aseptic conditions in a container or other suitable vessel, and perfused with a cell collection composition, or a standard perfusion solution (e.g., a normal saline solution such as phosphate buffered saline ("PBS") with or without an anticoagulant (e.g., heparin, warfarin sodium, coumarin, bishydroxycoumarin), and/or with or without an antimicrobial agent (e.g., ⁇ - mercaptoethanol (0.1 mM); antibiotics such as streptomycin (e.g., at 40-100 ⁇ g/ml), penicillin (e.g., at 40 U/ml), amphotericin B (e.g., at 0.5 ⁇ g/ml).
  • PBS phosphate buffered saline
  • the isolated placental cells banked in accordance with the methods described herein may be hematopoietic cells, natural killer cells isolated from placental perfusate, placental intermediate natural killer (Pi K) cells, placenta-derived adherent cells, or amnion-derived adherent cells.
  • the placenta-derived adherent cells express one or more genes at a level at least two-fold higher than bone marrow-derived mesenchymal stem cells, wherein said one or more genes are one or more of LOVL2, ST3GAL6, ST6GALNAC5, and/or SLC12A8.
  • the placenta-derived adherent cells express one or more genes at a level at least two-fold higher than bone marrow-derived mesenchymal stem cells, wherein said one or more genes are one or more of CPA4, TCF21, VTN, B4GALT6, FLJ 10781, and/or NUAK1.
  • the placenta-derived adherent cells are grown in culture, e.g., the placenta-derived adherent cells have adhered to a tissue culture substrate.
  • the cells are cultured in the presence of serum.
  • the cells are cultured in the absence of serum.
  • the placenta-derived adherent cells are substantially fetal in origin, e.g., the placenta-derived adherent cells are about or at least 90%, 95%, 99% or 100% fetal in origin.
  • the placenta-derived adherent cells are of mixed origin, e.g., the placenta-derived adherent cells are both fetal and maternal in origin.
  • the placenta-derived adherent cells have been passaged prior to isolation of exosomes from the placenta-derived adherent cells.
  • the placenta-derived adherent cells have been passaged about or at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, or 20 times.
  • the placenta-derived adherent cells have been cultured in vitro for 6 passages.
  • the isolated placental cells banked in accordance with the methods described herein may be natural killer (NK) cells.
  • NK cells are isolated from placental perfusate.
  • PiNK cells express one or more of the microRNAs hsa-miR- 100, hsa-miR-127, hsa-miR-211, hsa-miR-302c, hsa-miR-326, hsa-miR-337, hsa-miR-497, hsa- miR-512-3p, hsa-miR-515-5p, hsa-miR-517b, hsa-miR-517c, hsa-miR-518a, hsa-miR-518e, hsa- miR-519d, hsa-miR-520g, hsa-miR-520h, hsa-miR-564, hsa-miR-566, hsa-miR-618, and/or hsa- miR-99a at
  • Amnion derived adherent cells, and isolated populations of cells comprising the amnion derived adherent cells, provided herein can be produced by, e.g., digestion of amnion tissue followed by selection for adherent cells.
  • isolated amnion derived adherent cells can be produced by (1) digesting amnion tissue with a first enzyme to dissociate cells from the epithelial layer of the amnion from cells from the mesenchymal layer of the amnion; (2) subsequently digesting the mesenchymal layer of the amnion with a second enzyme to form a single-cell suspension; (3) culturing cells in said single-cell suspension on a tissue culture surface, e.g., tissue culture plastic; and (4) selecting cells that adhere to said surface after a change of medium, thereby producing an isolated population of cells comprising amnion derived adherent cells.
  • tissue culture surface e.g., tissue culture plastic
  • collagenase In a more specific embodiment, said collagenase is used at a concentration between 50 and 500 U/L in 5 mL per gram of amnion tissue to be digested. In another more specific embodiment, said digesting with collagenase is allowed to proceed for about 45-60 minutes at 37°C. In another specific embodiment, the single-cell suspension formed after digestion with collagenase is filtered through, e.g., a 75 ⁇ - 150 ⁇ filter between step (2) and step (3). In another specific embodiment, said first enzyme is trypsin, and said second enzyme is collagenase.
  • the cells adhere to a cell culture surface, e.g., to tissue culture plastic.
  • the OCT-4 " cell is VEGFR1/Flt-1 + (vascular endothelial growth factor receptor 1) and/or VEGFR2/KDR + (vascular endothelial growth factor receptor 2), as determined by immunolocalization.
  • the OCT-4 " amnion derived adherent cell, or a population of OCT-4 " amnion derived adherent cells expresses at least 2 log less PCR-amplified mRNA for OCT-4 at, e.g., 20 cycles, than an NTERA-2 cell, or population of NTERA-2 cells having an equivalent number of cells and RNA amplification cycles.
  • the cell is OCT-4 " , O49 , CD90 + , CD105 + , and CD117 " , as determined by immunolocalization or flow cytometry, and SOX2 , as determined by RT-PCR, e.g., for 30 cycles.
  • the cell is OCT-4 " as determined by RT-PCR, CD49 , CD90 + , and HLA-G " as determined by flow cytometry.
  • Placenta donors are recruited from expectant mothers that are enrolled in private umbilical cord blood banking programs and provide informed consent permitting the use of the exsanguinated placenta following recovery of cord blood for research purposes.
  • Donor data may be confidential.
  • the donor may be a customer of the bank, or the placenta may be donated to the bank by an individual who is not a customer of the bank. These donors also permit use of blinded data generated from the normal processing of their umbilical cord blood specimens for
  • cryopreservation This allows comparison between the composition of the collected cord blood and the effluent perfusate recovered using the experimental method described below.

Abstract

Provided herein are methods of banking placental tissue and cells derived from the same.

Description

METHODS OF BANKING PLACENTAL TISSUE AND CELLS
[0001] This application claims the benefit of U.S. Provisional Application No. 62/134,943, filed March 18, 2015, which is hereby incorporated by reference in its entirety.
1. FIELD
[0002] The disclosure herein relates to methods of banking placental tissue.
2. BACKGROUND
[0003] The preservation and subsequent use of tissues and cells is of great importance to the medical and scientific fields. There remains a need in the field, however, for improved methods of banking and preserving tissue, including placental tissue.
3. SUMMARY
[0004] In one aspect, provided herein are methods of banking placental tissue in a tissue bank.
[0005] In one embodiment, the methods of banking placental tissue provided herein comprise: (a) receiving a placenta or a portion thereof, (b) processing the placenta, e.g., to clean the placenta and/or isolate placental tissue, and (c) banking the placental tissue. In certain embodiments, the banked placental tissue comprises one or more of (i) whole placenta; (ii) amniotic membrane; (iii) chorionic membrane; (iv) whole umbilical cord blood; (v) umbilical cord vessels; (vi) umbilical cord membrane; (vii) placental vessels; (viii) platelet-rich plasma from placental blood or umbilical cord blood; (ix) isolated placental cells; (x) placental perfusate; (xi) human placenta-derived stem cells (HPDSC) and/or (xii) cells isolated from placental perfusate. In a specific embodiment, the banked placental tissue comprises a whole placenta. In another specific embodiment, the banked placental tissue comprises human placenta-derived stem cells (HPDSC). In another specific embodiment, the banked placental tissue comprises cells isolated from placental perfusate.
[0006] In another embodiment, the methods of banking placental tissue provided herein comprise: (a) receiving a placenta or a portion thereof from a customer, (b) obtaining placental blood and/or umbilical cord from said placenta, (c) providing the customer with a selection of placental tissue from said placenta or portion thereof to bank, (d) receiving an election from the customer as to which placental tissue to bank, (e) processing the placenta to isolate the elected tissue(s), and (f) banking the placental tissue. In certain embodiments, the banked placental tissue comprises one or more of (i) whole placenta; (ii) amniotic membrane; (iii) chorionic membrane; (iv) whole umbilical cord blood; (v) umbilical cord vessels; (vi) umbilical cord membrane; (vii) placental vessels; (viii) platelet-rich plasma from placental blood or umbilical cord blood; (ix) isolated placental cells; (x) placental perfusate; (xi) human placenta-derived stem cells (HPDSC) and/or (xii) cells isolated from placental perfusate. In a specific
embodiment, the banked placental tissue comprises a whole placenta. In another specific embodiment, the banked placental tissue comprises human placenta-derived stem cells
(HPDSC). In another specific embodiment, the banked placental tissue comprises cells isolated from placental perfusate.
[0007] In one embodiment of the methods described herein, the placental tissue that is banked comprises whole placenta, e.g., intact placenta. In a specific embodiment, said whole placenta is decellularized. Methods of decellularization are well known in the art and include, without limitation, enzymatic digestion (e.g., trypsinization), use of detergents, and freeze-thaw methods. In one embodiment, the placenta has been exsanguinated prior to banking. In one embodiment, placental perfusate is collected from a placenta that has been exsanguinated prior to banking. In another embodiment, said placental perfusate has been collected from a placenta that has been exsanguinated but not perfused to remove residual blood. In another embodiment, said placental perfusate has been collected from a placenta that has not been exsanguinated.
[0008] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is an exsanguinated placenta. In one embodiment of the methods described herein, the exsanguinated placenta is additionally banked with blood obtained from the exsanguinated placenta, e.g., whole cord blood.
[0009] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is a perfused placenta. In one embodiment of the methods described herein, the perfused placenta is additionally banked with placental perfusate obtained from the placenta.
[0010] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is an exsanguinated and perfused placenta. In another embodiment, an exsanguinated and perfused placenta is banked using the methods described herein, and blood obtained from the exsanguinated placenta, e.g., whole cord blood, and/or placental perfusate obtained from the placenta is/are banked with the exsanguinated and perfused placenta. Methods of isolating blood/cells from an exsanguinated and/or perfused placenta are known in the art and described in Section 4.2.
[0011] In one embodiment, the placental tissue banked in accordance with the methods described herein is placental perfusate. In a specific embodiment, the placental tissue banked in accordance with the methods described herein is total nucleated cells isolated from placental perfusate (i.e., HPDSC). In another specific embodiment, placental perfusate and/or total nucleated cells isolated from placental perfusate are banked along with one or more of whole umbilical cord blood, amniotic membrane, chorionic membrane, umbilical cord vessels, umbilical cord membrane, placental vessels, platelet-rich plasma from placental blood or umbilical cord blood, and/or isolated placental cells (see Section 4.3).
[0012] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is whole umbilical cord blood. In a specific embodiment, said whole umbilical cord blood is banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, chorionic membrane, umbilical cord, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), umbilical cord membrane, placental vessels, platelet-rich plasma from placental blood, and/or isolated cells from the placenta (see Section 4.3). In another specific embodiment, said whole umbilical cord blood is banked with human placenta-derived stem cells (HPDSC).
[0013] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is amniotic membrane. In a specific embodiment, said amniotic membrane is decellularized. In another specific embodiment, said amniotic membrane is banked with one or more of placental perfusate, cells isolated from placental perfusate, chorionic membrane, umbilical cord, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), umbilical cord membrane, placental vessels, platelet-rich plasma from placental blood or umbilical cord blood, and/or isolated cells from the placenta (see Section 4.3). In another specific embodiment, said amniotic membrane is banked with human placenta-derived stem cells (HPDSC).
[0014] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is chorionic membrane In a specific embodiment, said chorionic membrane is decellularized. In another specific embodiment, said chorionic membrane is banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, umbilical cord, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), umbilical cord membrane, placental vessels, platelet-rich plasma from placental blood or umbilical cord blood, and isolated cells from the placenta (see Section 4.3). In another specific embodiment, said chorionic membrane is banked with human placenta-derived stem cells (HPDSC).
[0015] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is umbilical cord. In a specific embodiment, said umbilical cord is
decellularized. In another specific embodiment, said umbilical cord is banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), umbilical cord membrane, placental vessels, platelet-rich plasma from placental blood or umbilical cord blood, and isolated cells from the placenta (see Section 4.3). In another specific embodiment, said umbilical cord is banked with human placenta-derived stem cells (HPDSC).
[0016] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is one or more umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein). In a specific embodiment, said umbilical cord vessels are decellularized. In another specific embodiment, said umbilical cord vessels are banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, chorionic membrane, umbilical cord, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord membrane, placental vessels, platelet-rich plasma from placental blood or umbilical cord blood, and/or isolated cells from the placenta (see Section 4.3). In another specific embodiment, said umbilical cord vessels are banked with human placenta-derived stem cells (HPDSC).
[0017] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is umbilical cord membrane. In a specific embodiment, said umbilical cord membrane is decellularized. In another specific embodiment, said umbilical cord membrane is banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, chorionic membrane, umbilical cord, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), placental vessels, platelet-rich plasma from placental blood or umbilical cord blood, and/or isolated cells from the placenta (see Section 4.3). In another specific embodiment, said umbilical cord membrane is banked with human placenta-derived stem cells (HPDSC).
[0018] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is one or more placental vessels. In a specific embodiment, said placental vessels are banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, chorionic membrane, umbilical cord, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), umbilical cord membrane, platelet-rich plasma from placental blood or umbilical cord blood, and/or isolated cells from the placenta (see Section 4.3). In another specific embodiment, said placental vessels are banked with human placenta-derived stem cells (HPDSC).
[0019] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is platelet-rich plasma from placental blood and/or platelet-rich plasma from umbilical cord blood. In a specific embodiment, said platelet-rich plasma from placental blood or platelet-rich plasma from umbilical cord blood is banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, chorionic membrane, umbilical cord, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), umbilical cord membrane, placental vessels, and/or isolated cells from the placenta (see Section 4.3). In another specific embodiment, said platelet-rich plasma from placental blood or platelet-rich plasma from umbilical cord blood is banked with human placenta-derived stem cells (HPDSC).
[0020] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is isolated placental cells. See Section 4.3. Isolated placental stem cells that can be banked in accordance with the methods described herein include CD34+ hematopoietic stem cells, e.g., CD34+ hematopoietic stem cells isolated from placental perfusate (see Section 4.3.1.1); HPDSC; natural killer cells, e.g., natural killer cells isolated from placental perfusate (see Section 4.3.1.3); CD10+, CD34", CD105+ CD200+ tissue culture plastic adherent cells (see Section 4.3.1.2); and amni on-derived adherent cells (see Section 4.3.1.4). In a specific embodiment, said isolated placental cells are banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, chorionic membrane, umbilical cord, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), umbilical cord membrane, placental vessels, and/or platelet-rich plasma from placental blood or umbilical cord blood.
[0021] In one embodiment, the banked placental tissue comprises or consists of placental perfusate. In a specific embodiment, the banked placental tissue comprises or consists of placental perfusate and umbilical cord blood. In another embodiment, the banked placental tissue comprises or consists of human placenta-derived stem cells (HPDSC). In a specific embodiment, the banked placental tissue comprises or consists of human placenta-derived stem cells (HPDSC) and umbilical cord blood. In another embodiment, the banked placental tissue comprises or consists of umbilical cord vessels. In a specific embodiment, the banked placental tissue comprises or consists of umbilical cord vessels and umbilical cord blood. In another embodiment, the banked placental tissue comprises or consists of umbilical cord. In a specific embodiment, the banked placental tissue comprises or consists of umbilical cord and umbilical cord blood. In another embodiment, the banked placental tissue comprises or consists of an intact placenta. In a specific embodiment, the banked placental tissue comprises or consists of an intact placenta and umbilical cord blood. In another embodiment, the banked placental tissue comprises or consists of umbilical cord blood and placental blood. In a specific embodiment, the banked placental tissue comprises or consists of umbilical cord blood, placental blood, and one or more placental tissues selected from the group consisting of (i) whole placenta; (ii) amniotic membrane; (iii) chorionic membrane; (iv) whole umbilical cord blood; (v) umbilical cord vessels; (vi) umbilical cord membrane; (vii) placental vessels; (viii) isolated placental cells; (ix) placental perfusate; (x) human placenta-derived stem cells (HPDSC) and/or (xi) cells isolated from placental perfusate.
[0022] In one embodiment of the methods described herein, the placental tissue to be banked is determined by a customer of the bank in which the placental tissue is banked, i.e., the customer elects what placental tissue is to be banked, e.g., from a list of possible tissues that is provided by the bank.
[0023] In a specific embodiment, the methods described herein comprise banking of (i) at least one unit of placental blood or umbilical cord blood and (ii) at least one unit of placental perfusate or total nucleated cells collected from placental perfusate, wherein said placental or umbilical cord blood and said placental perfusate or total nucleated cells collected from said placental perfusate are isolated from the same placenta.
[0024] In certain embodiments, the placental tissue banked in accordance with the methods described herein is cryopreserved. In a specific embodiment, the placental tissue is
cryopreserved by the placental tissue bank.
[0025] In certain embodiments, the placental tissue banked in accordance with the methods described herein is allogeneic to the individual that has elected to bank the placental tissue, e.g., the customer of the placental tissue bank. In certain embodiments, the placental tissue banked in accordance with the methods described herein is autologous to the individual that has elected to bank the placental tissue, e.g., the customer of the placental tissue bank.
[0026] In certain embodiments, the methods described herein comprise assigning the placental tissue to be banked an identifier based on the source of the placenta. In another embodiment, the methods described herein comprise assigning the placenta that is the source of placental tissue to be banked a first identifier, and assigning each placental tissue to be banked from the placenta a second tissue-specific identifier. In certain embodiments, the second tissue-specific identifier incorporates said first identifier (for example, the placenta can be assigned the identifier XYZ, and amnion from the placenta can be assigned the identifier XYZ 123). In certain embodiments, the placental tissue bank in which the placental tissue is banked comprises a system for storing and retrieving the identifiers, e.g., the first and second identifiers.
[0027] In another aspect, provided herein are methods of providing placental tissue to a customer of the placental tissue bank. In a specific embodiment, a method of providing placental tissue to a customer of the placental tissue bank comprises: (a) providing the customer a selection of placental tissue from said placenta or portion thereof to bank, (b) receiving an election from the customer as to which placental tissue to bank; (c) processing the placenta to isolate the elected tissue(s); and (d) banking the tissues at the direction of said customer, thereby providing placental tissue to a customer.
4. DETAILED DESCRIPTION
4.1. Methods of Banking Placental Tissue
[0028] The methods described herein are directed to the banking of placental tissue in a tissue bank. As used herein, "placental tissue" can include whole placenta, placental perfusate, cells isolated from placental perfusate, amniotic membrane, chorionic membrane, umbilical cord, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), umbilical cord membrane, placental vessels, platelet-rich plasma from placental blood or umbilical cord blood, and/or isolated cells from the placenta, e.g., cells described in Section 4.3.
[0029] A "bank," e.g., a "placental tissue bank," as described herein, refers to an entity comprising one or more persons that is responsible for the storage, management, and retrieval of placental tissue and/or cells derived therefrom. "Banking," as described herein, refers to the essential functions of the bank, including but not limited to the reception, storage (e.g., cryopreservation), recordation, and retrieval of placental tissue and/or cells derived therefrom. The bank additionally receives elections from customers as to which one or more placental tissue and/or cells derived therefrom are to be stored in the bank, and assigns to said one or more tissues and/or cells derived therefrom an identifier according to the methods described herein. Said identifier allows the bank to monitor and retrieve said one or more placental tissue and/or cells derived therefrom. A "customer," as referred to herein, is one or more individuals using the services of a bank described herein for the banking of one or more placental tissue. After banking said tissue, a customer may subsequently request said tissue using the one or more identifiers assigned by the bank (see Section 4.1.3).
[0030] "Human placenta-derived stem cells" (HPDSC), as used herein, mean the cells isolated from placental perfusate, e.g., by centrifugation. Typically HPDSC are a heterogeneous population of nucleated placental cells from perfusate, from which enucleated cells have been removed. HPDSC typically include a sub-population of CD34+ cells, e.g., hematopoietic stem cells, and/or a population of tissue culture plastic-adherent CD34" placental stem cells, e.g., as described in U.S. Patent Nos. 7,468,276; 8,057,788 and 8,202,703, the disclosures of which are hereby incorporated by reference in their entireties.
[0031] In one aspect, provided herein are methods of banking placental tissue in a tissue bank.
[0032] In one embodiment, the methods of banking placental tissue provided herein comprise: (a) receiving a placenta or a portion thereof, (b) processing the placenta, e.g., to clean the placenta and/or isolate placental tissue, and (c) banking the placental tissue. In certain embodiments, the banked placental tissue comprises one or more of (i) whole placenta; (ii) amniotic membrane; (iii) chorionic membrane; (iv) whole umbilical cord blood; (v) umbilical cord vessels; (vi) umbilical cord membrane; (vii) placental vessels; (viii) platelet-rich plasma from placental blood or umbilical cord blood; (ix) isolated placental cells; (x) placental perfusate; (xi) human placenta-derived stem cells (HPDSC) and/or (xii) cells isolated from placental perfusate. In a specific embodiment, the banked placental tissue comprises a whole placenta. In another specific embodiment, the banked placental tissue comprises human placenta-derived stem cells (HPDSC). In another specific embodiment, the banked placental tissue comprises cells isolated from placental perfusate.
[0033] In another embodiment, the methods of banking placental tissue provided herein comprise: (a) receiving a placenta or a portion thereof from a customer, (b) obtaining placental blood and/or umbilical cord from said placenta, (c) providing the customer with a selection of placental tissue from said placenta or portion thereof to bank, (d) receiving an election from the customer as to which placental tissue to bank, (e) processing the placenta to isolate the elected tissue(s), and (f) banking the placental tissue. In certain embodiments, the banked placental tissue comprises one or more of (i) whole placenta; (ii) amniotic membrane; (iii) chorionic membrane; (iv) whole umbilical cord blood; (v) umbilical cord vessels; (vi) umbilical cord membrane; (vii) placental vessels; (viii) platelet-rich plasma from placental blood or umbilical cord blood; (ix) isolated placental cells; (x) placental perfusate; (xi) human placenta-derived stem cells (HPDSC) and/or (xii) cells isolated from placental perfusate. In a specific
embodiment, the banked placental tissue comprises a whole placenta. In another specific embodiment, the banked placental tissue comprises human placenta-derived stem cells
(HPDSC). In another specific embodiment, the banked placental tissue comprises cells isolated from placental perfusate.
[0034] In one embodiment of the methods described herein, the placental tissue that is banked comprises whole placenta, e.g., intact placenta. In a specific embodiment, said whole placenta is decellularized. Methods of decellularization are well known in the art and include, without limitation, enzymatic digestion (e.g., trypsinization), use of detergents, and freeze-thaw methods. In one embodiment, the placenta has been exsanguinated prior to banking. In one embodiment, placental perfusate is collected from a placenta that has been exsanguinated prior to banking. In another embodiment, said placental perfusate has been collected from a placenta that has been exsanguinated but not perfused to remove residual blood. In another embodiment, said placental perfusate has been collected from a placenta that has not been exsanguinated. [0035] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is an exsanguinated placenta. In one embodiment of the methods described herein, the exsanguinated placenta is additionally banked with blood obtained from the exsanguinated placenta, e.g., whole cord blood.
[0036] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is a perfused placenta. In one embodiment of the methods described herein, the perfused placenta is additionally banked with placental perfusate obtained from the placenta.
[0037] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is an exsanguinated and perfused placenta. In another embodiment, an exsanguinated and perfused placenta is banked using the methods described herein, and blood obtained from the exsanguinated placenta, e.g., whole cord blood, and/or placental perfusate obtained from the placenta is/are banked with the exsanguinated and perfused placenta. Methods of isolating blood/cells from an exsanguinated and/or perfused placenta are known in the art and described in Section 4.2.
[0038] In one embodiment, the placental tissue banked in accordance with the methods described herein is placental perfusate. In a specific embodiment, the placental tissue banked in accordance with the methods described herein is total nucleated cells isolated from placental perfusate. In another specific embodiment, placental perfusate and/or total nucleated cells isolated from placental perfusate are banked along with one or more of whole umbilical cord blood, amniotic membrane, chorionic membrane, umbilical cord vessels, umbilical cord membrane, placental vessels, platelet-rich plasma from placental blood or umbilical cord blood, and/or isolated placental cells (see Section 4.3).
[0039] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is whole umbilical cord blood. In a specific embodiment, said whole umbilical cord blood is banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, chorionic membrane, umbilical cord, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), umbilical cord membrane, placental vessels, platelet-rich plasma from placental blood, and/or isolated cells from the placenta (see Section 4.3).
[0040] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is amniotic membrane. In a specific embodiment, said amniotic membrane is decellularized. In another specific embodiment, said amniotic membrane is banked with one or more of placental perfusate, cells isolated from placental perfusate, chorionic membrane, umbilical cord, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), umbilical cord membrane, placental vessels, platelet-rich plasma from placental blood or umbilical cord blood, and/or isolated cells from the placenta (see Section 4.3).
[0041] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is chorionic membrane In a specific embodiment, said chorionic membrane is decellularized. In another specific embodiment, said chorionic membrane is banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, umbilical cord, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), umbilical cord membrane, placental vessels, platelet-rich plasma from placental blood or umbilical cord blood, and isolated cells from the placenta (see Section 4.3).
[0042] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is umbilical cord. In a specific embodiment, said umbilical cord is
decellularized. In another specific embodiment, said umbilical cord is banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), umbilical cord membrane, placental vessels, platelet-rich plasma from placental blood or umbilical cord blood, and isolated cells from the placenta (see Section 4.3).
[0043] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is one or more umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein). In a specific embodiment, said umbilical cord vessels are decellularized. In another specific embodiment, said umbilical cord vessels are banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, chorionic membrane, umbilical cord, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord membrane, placental vessels, platelet-rich plasma from placental blood or umbilical cord blood, and/or isolated cells from the placenta (see Section 4.3). [0044] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is umbilical cord membrane. In a specific embodiment, said umbilical cord membrane is decellularized. In another specific embodiment, said umbilical cord membrane is banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, chorionic membrane, umbilical cord, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), placental vessels, platelet-rich plasma from placental blood or umbilical cord blood, and/or isolated cells from the placenta (see Section 4.3).
[0045] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is one or more placental vessels. In a specific embodiment, said placental vessels are banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, chorionic membrane, umbilical cord, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), umbilical cord membrane, platelet-rich plasma from placental blood or umbilical cord blood, and/or isolated cells from the placenta (see Section 4.3).
[0046] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is platelet-rich plasma from placental blood and/or platelet-rich plasma from umbilical cord blood. In a specific embodiment, said platelet-rich plasma from placental blood or platelet-rich plasma from umbilical cord blood is banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, chorionic membrane, umbilical cord, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), umbilical cord membrane, placental vessels, and/or isolated cells from the placenta (see Section 4.3).
[0047] In one embodiment, the placental tissue that is banked in accordance with the methods described herein is isolated placental cells. See Section 4.3. Isolated placental stem cells that can be banked in accordance with the methods described herein include CD34+ hematopoietic stem cells, e.g., CD34+ hematopoietic stem cells isolated from placental perfusate (see Section 4.3.1.1); HPDSC; natural killer cells, e.g., natural killer cells isolated from placental perfusate (see Section 4.3.1.3); CD10+, CD34-, CD105+ CD200+ tissue culture plastic adherent cells (see Section 4.3.1.2); and amni on-derived adherent cells (see Section 4.3.1.4). In a specific embodiment, said isolated placental cells are banked with one or more of placental perfusate, cells isolated from placental perfusate, amniotic membrane, chorionic membrane, umbilical cord, umbilical cord blood, cells isolated from umbilical cord blood, umbilical cord vessels (e.g., umbilical cord arteries or the umbilical cord vein), umbilical cord membrane, placental vessels, and/or platelet-rich plasma from placental blood or umbilical cord blood.
[0048] In one embodiment, the banked placental tissue comprises or consists of placental perfusate. In a specific embodiment, the banked placental tissue comprises or consists of placental perfusate and umbilical cord blood. In another embodiment, the banked placental tissue comprises or consists of human placenta-derived stem cells (HPDSC). In a specific embodiment, the banked placental tissue comprises or consists of human placenta-derived stem cells (HPDSC) and umbilical cord blood. In another embodiment, the banked placental tissue comprises or consists of umbilical cord vessels. In a specific embodiment, the banked placental tissue comprises or consists of umbilical cord vessels and umbilical cord blood. In another embodiment, the banked placental tissue comprises or consists of umbilical cord. In a specific embodiment, the banked placental tissue comprises or consists of umbilical cord and umbilical cord blood. In another embodiment, the banked placental tissue comprises or consists of an intact placenta. In a specific embodiment, the banked placental tissue comprises or consists of an intact placenta and umbilical cord blood. In another embodiment, the banked placental tissue comprises or consists of umbilical cord blood and placental blood. In a specific embodiment, the banked placental tissue comprises or consists of umbilical cord blood, placental blood, and one or more placental tissues selected from the group consisting of (i) whole placenta; (ii) amniotic membrane; (iii) chorionic membrane; (iv) whole umbilical cord blood; (v) umbilical cord vessels; (vi) umbilical cord membrane; (vii) placental vessels; (viii) isolated placental cells; (ix) placental perfusate; (x) human placenta-derived stem cells (HPDSC) and/or (xi) cells isolated from placental perfusate.
[0049] In one embodiment of the methods described herein, the placental tissue to be banked is determined by a customer of the bank in which the placental tissue is banked, i.e., the customer elects what placental tissue is to be banked, e.g., from a list of possible tissues that is provided by the bank.
[0050] In a specific embodiment, the methods described herein comprise banking of (i) at least one unit of placental blood or umbilical cord blood and (ii) at least one unit of placental perfusate or total nucleated cells collected from placental perfusate, wherein said placental or umbilical cord blood and said placental perfusate or total nucleated cells collected from said placental perfusate are isolated from the same placenta.
[0051] In certain embodiments, the placental tissue banked in accordance with the methods described herein is cryopreserved. In a specific embodiment, the placental tissue is
cryopreserved by the placental tissue bank.
[0052] In certain embodiments, the placental tissue banked in accordance with the methods described herein is allogeneic to the individual that has elected to bank the placental tissue, e.g., the customer of the placental tissue bank. In certain embodiments, the placental tissue banked in accordance with the methods described herein is autologous to the individual that has elected to bank the placental tissue, e.g., the customer of the placental tissue bank.
[0053] In certain embodiments, the methods described herein comprise assigning the placental tissue to be banked an identifier based on the source of the placenta. In another embodiment, the methods described herein comprise assigning the placenta that is the source of placental tissue to be banked a first identifier, and assigning each placental tissue to be banked from the placenta a second tissue-specific identifier. In certain embodiments, the second tissue-specific identifier incorporates said first identifier (for example, the placenta can be assigned the identifier XYZ, and amnion from the placenta can be assigned the identifier XYZ 123). In certain embodiments, the placental tissue bank in which the placental tissue is banked comprises a system for storing and retrieving the identifiers, e.g., the first and second identifiers.
[0054] In another aspect, provided herein are methods of providing placental tissue to a customer of the placental tissue bank. In a specific embodiment, a method of providing placental tissue to a customer of the placental tissue bank comprises: (a) providing the customer a selection of placental tissue from said placenta or portion thereof to bank, (b) receiving an election from the customer as to which placental tissue to bank; (c) processing the placenta to isolate the elected tissue(s); and (d) banking the tissues at the direction of said customer, thereby providing placental tissue to a customer.
4.1.1. Preservation of Placental Tissue
[0055] The methods described herein comprise banking of placental tissue. In certain embodiments, the placental tissue is frozen (e.g., cryopreserved) prior to or at the time of banking. Method of freezing placental tissue that can be used in accordance with the methods of banking described herein include freezing of the tissue in the presence of dry ice, in a freezer (e.g., a freezer at -80°C), and in liquid nitrogen. In a specific embodiment, said freezing (e.g., cryopreserving) is performed by the placental tissue bank. In certain embodiments, the placental tissue banked in accordance with the methods described herein is not frozen (e.g., not cryopreserved) as part of the banking method.
4.1.2. Origin of Placental Tissue
[0056] The methods described herein may be used for the banking of placental tissue derived from one or more sources (e.g., one or more placentas). In a specific embodiment, said placental tissue is autologous to a customer of the placental tissue bank that banks the placental tissue. In another embodiment, said placental tissue is heterologous to a customer of the placental tissue bank that banks the placental tissue. The source of the placental tissue may be any placental animal (e.g., a primate, for example a human). In a specific embodiment, said placental tissue is derived from a human.
4.1.3. Placental Tissue Identification and Retrieval
[0057] The methods described herein comprise the banking of placental tissue, and may additionally comprise the assignment of identifiers to the placental tissue to allow for subsequent monitoring and retrieval of the placental tissue after banking. In one embodiment, the placental tissue is assigned an identifier based on the region of the placenta from which said placental tissue is derived. In another embodiment, the placenta that is the source of placental tissue to be banked is assigned a first identifier, and each placental tissue to be obtained from the placenta (e.g., amnion, chorion, etc.) is assigned a second, tissue-specific, identifier specific for the particular tissue. In a specific embodiment, said second identifier incorporates said first identifier into a single identifier. In another embodiment, the placental tissue bank that banks the placental tissue comprises a system for storing and subsequently retrieving said identifiers, e.g., said first and second identifiers.
[0058] In another specific embodiment of the methods described herein, placental tissue is banked in a placental tissue bank, wherein the method comprises the steps of (a) receiving a placenta or portion thereof, (b) processing the placenta to isolate the placental tissue as described herein, and (c) subsequently banking the placental tissue, wherein prior to step (a) or step (b), a customer elects which placental tissue from said placenta to bank. In another specific embodiment, placental tissue is banked in a placental tissue bank according to a method comprising (a) receiving an election from a customer of the placental tissue bank as to which placental tissue(s) to bank; (b) receiving a placenta or portion thereof; (c) processing the placenta to isolate the placental tissue as described herein; and (d) subsequently banking the elected placental tissue. In another specific embodiment of the methods described herein, the one or more placental tissue elected by a customer are assigned a third identifier to indicate election. In yet another embodiment of the methods described herein, the one or more placental tissue not elected by the customer are assigned a fourth identifier to indicate non-election. For example, prior to the bank receiving the placenta, a customer may wish to bank a specific placental tissue (e.g., amniotic membrane). This elected tissue is assigned a unique fourth identifier, while each of the non-elected placental tissues is assigned a different third identifier to indicate non-election. In one embodiment, the one or more placental tissues that are assigned unique identifiers (e.g., a first, second, third, and/or fourth identifier) are recorded on a computer or in paper records.
[0059] In one embodiment, the unique identifiers described herein may be public or private identifiers. In another embodiment, a customer receives a unique identifier. In a specific embodiment, said identifier assigned to the customer is private. In another embodiment, one or more of the unique identifiers described herein is public. Said public identifier may be used to indicate elected or non-elected placental tissues in the records of one bank that may be viewed by another bank.
[0060] In certain embodiments of the methods described herein, a customer may elect to bank an intact placenta or one or more of several types of placental tissue. In a specific embodiment, provided herein is a method of providing placental tissue to a customer comprising providing the customer a selection of placental tissue from said placenta or portion thereof to a bank, wherein the tissues comprise one or both of whole placental perfusate and/or cells isolated from placental perfusate. In additional embodiments of the methods described herein, said tissues provided to a bank may comprise (1) one or both of whole placenta perfusate and/or cells isolated from placental perfusate and (2) additionally one or more of amniotic membrane, chorionic
membrane, whole umbilical cord blood, umbilical cord vessels, umbilical cord membrane, placental vessels, platelet-rich plasma from placental blood or umbilical cord blood, and isolated placental cells. In other embodiments, the methods described herein comprise providing placental tissue to a customer comprising providing the customer a selection of one or more placental tissue as described herein, receiving an election from the customer as to which placental tissue to bank, processing the placenta to isolate the elected tissues as described herein, and banking the tissues at the direction of said customer. In specific embodiments, the methods described herein may comprise additionally providing umbilical cord blood and/or placental blood to a customer. In a specific embodiment, said umbilical cord blood and/or placental blood are autologous to the customer. In another embodiment, said umbilical cord blood and/or placental blood are heterologous to the customer.
[0061] In one embodiment of the methods described herein, placental tissue banking comprises receiving from a customer placental tissue (e.g., an intact placenta or a portion thereof), obtaining placental blood and/or umbilical cord blood from said placental tissue, providing the customer with a selection of placental tissue from said placenta or portion thereof to bank, receiving an election from the customer as to which one or more placental tissue to bank, processing the placenta to isolate the elected one or more placental tissue, and banking the placental tissue, wherein said placental tissue further comprise whole placenta, amniotic membrane, chorionic membrane, whole umbilical cord blood, umbilical cord vessels, umbilical cord membrane, placental vessels, platelet-rich plasma from placental blood or umbilical cord blood, isolated placental cells, placental perfusate, or cells isolated from placental perfusate, and optionally baking said umbilical cord blood or said placental blood. In another embodiment, the customer selects the tissue for banking to be one or more of whole placenta, amniotic membrane, chorionic membrane, whole umbilical cord blood, umbilical cord vessels, umbilical cord membrane, placental vessels, and additionally comprising providing the customer with the option of decellularizing said placental tissue.
4.2. Placental Processing
[0062] In accordance with the methods described herein, an intact placenta may be processed prior to its banking. In one embodiment, the processed placental tissue may be an intact placenta, e.g., an intact placenta that has been exsanguinated or an intact placenta that has been
exsanguinated and perfused (for example, perfused as described in Section 4.2.1, infra). In another embodiment, the processed placental tissue may be placental perfusate {e.g., the placental perfusate isolated as described in Section 4.2.1). In another embodiment, the placental tissue consists of the total nucleated cells isolated from placental perfusate {e.g., total nucleated cells isolated from a placenta perfused according to the methods described in Section 4.2.1).
[0063] In a particular embodiment, for example, an intact placenta is drained of cord blood, further processed, e.g., perfused, to remove residual blood, and perfused to obtain placental perfusate, as for example, described in Section 4.2.1, infra, prior to banking. In another particular embodiment, an intact placenta is drained of cord blood and further processed, e.g., perfused, to remove residual blood but is not perfused to obtain placental perfusate prior to banking. In yet another embodiment, an intact placenta is drained of cord blood but is not further processed to remove residual blood or perfused to obtain placental perfusate prior to banking.
[0064] In one embodiment, a method of banking placental tissue comprises receiving a placenta or a portion thereof, processing said placenta or portion thereof to isolate the placental tissue, and banking said placental tissue comprising placental perfusate or total nucleated cells isolated from said placental perfusate. Typically, placental perfusate from a single placental perfusion comprises about 100 million to about 500 million nucleated cells.
[0065] Generally, a human placenta is recovered shortly after its expulsion after birth. In one embodiment, the placenta is recovered from a patient after informed consent and after a complete medical history of the patient is taken and is associated with the placenta. In another
embodiment, the tracking of medical history continues after delivery.
[0066] In certain embodiments, the intact placenta is cleaned prior to further processing and handling. In one embodiment, the intact placenta is handled to separate its constituent tissues and cell types after cleaning. In one embodiment, placental tissues are dissected and isolated {e.g., the amniotic membrane, the chorionic membrane, umbilical cord vessels, umbilical cord membrane, and placental vessels are dissected and isolated) after cleaning the intact placenta. In another embodiment, cells associated with the placenta are isolated {e.g., umbilical cord blood is isolated) after cleaning the intact placenta. In a specific embodiment, the intact placenta is not dissected after cleaning. In specific embodiments, the intact placenta or placental tissues derived therefrom are cryopreserved according to the methods described herein (see Section 5.1.2).
4.2.1. Placental Perfusion
[0067] Methods of perfusing mammalian placentas and obtaining placental perfusate are disclosed, e.g., in Hariri, U.S. Patent Nos. 7,045,148 and 7,255,879, 8,057,788 and in U.S.
Application Publication Nos. 2009/0104164, 2007/0190042, the disclosures of which are hereby incorporated by reference herein in their entireties.
[0068] In certain embodiments, placental perfusate is collected before banking placental tissue and/or cells derived therefrom. In a particular embodiment, prior to recovery of perfusate, the umbilical cord blood and placental blood are removed. In certain embodiments, after delivery, the cord blood in the placenta is recovered. In a particular embodiment, prior to recovery of placental perfusate, the placenta is drained of cord blood and further processed, e.g., perfused, to remove residual blood. In another particular embodiment, prior to recovery of placental perfusate, the placenta is drained of cord blood, but is not further processed to remove residual blood. In yet another embodiment, prior to recovery of placental perfusate, the placenta is neither drained of cord blood nor processed to remove residual blood.
[0069] The placenta can be subjected to a conventional cord blood recovery process. Typically a needle or cannula is used, with the aid of gravity, to exsanguinate the placenta (see, e.g., Anderson, U.S. Patent No. 5,372,581; Hessel et al., U.S. Patent No. 5,415,665). The needle or cannula is usually placed in the umbilical vein and the placenta can be gently massaged to aid in draining cord blood from the placenta. Such cord blood recovery may be performed
commercially, e.g., LifeBank Inc., Cedar Knolls, N.J., ViaCord, Cord Blood Registry and CryoCell. In one embodiment, the placenta is gravity drained without further manipulation so as to minimize tissue disruption during cord blood recovery.
[0070] Typically, a placenta is transported from the delivery or birthing room to another location, e.g., a laboratory, for recovery of cord blood and collection of perfusate. The placenta can be transported in a sterile, thermally insulated transport device (maintaining the temperature of the placenta between 20-28°C), for example, by placing the placenta, in a sterile zip-lock plastic bag, which is then placed in an insulated container. In another embodiment, the placenta is transported in a cord blood collection kit substantially as described in U.S. Patent No.
7, 147,626, the disclosure of which is hereby incorporated by reference herein in its entirety. In one embodiment, the placenta is delivered to the laboratory four to twenty-four hours following delivery. In certain embodiments, the proximal umbilical cord is clamped, for example within 4- 5 cm (centimeter) of the insertion into the placental disc prior to cord blood recovery. In other embodiments, the proximal umbilical cord is clamped after cord blood recovery but prior to further processing of the placenta.
[0071] The placenta, prior to collection of the perfusate, can be stored under sterile conditions and at either room temperature or at a temperature of 5 to 25°C. The placenta may be stored for a period of longer than forty eight hours, or for a period of four to twenty -four hours prior to perfusing the placenta to remove any residual cord blood. The placenta can be stored in an anticoagulant solution at a temperature of 5°C to 25°C. Suitable anticoagulant solutions are well known in the art. For example, a solution of heparin or warfarin sodium can be used. In one embodiment, the anticoagulant solution comprises a solution of heparin (e.g., 1% w/v in 1 : 1000 solution). In some embodiments, the exsanguinated placenta is stored for no more than 36 hours before placental perfusate is collected.
[0072] Perfusate can be obtained by passage of perfusion solution, e.g., saline solution, culture medium or cell collection compositions described above, through the placental vasculature. In one embodiment, a mammalian placenta is perfused by passage of perfusion solution through either or both of the umbilical artery and umbilical vein. The flow of perfusion solution through the placenta may be accomplished using, e.g., gravity flow into the placenta. For example, the perfusion solution is forced through the placenta using a pump, e.g., a peristaltic pump. The umbilical vein can be, e.g., cannulated with a cannula, e.g., a TEFLON® or plastic cannula, that is connected to a sterile connection apparatus, such as sterile tubing. The sterile connection apparatus is connected to a perfusion manifold.
[0073] In preparation for perfusion, the placenta can be oriented in such a manner that the umbilical artery and umbilical vein are located at the highest point of the placenta. The placenta can be perfused by passage of a perfusion solution through the placental vasculature, or through the placental vasculature and surrounding tissue. In one embodiment, the umbilical artery and the umbilical vein are connected simultaneously to a pipette that is connected via a flexible connector to a reservoir of the perfusion solution. The perfusion solution is passed into the umbilical vein and artery. The perfusion solution exudes from and/or passes through the walls of the blood vessels into the surrounding tissues of the placenta, and is collected in a suitable open vessel from the surface of the placenta that was attached to the uterus of the mother during gestation. The perfusion solution may also be introduced through the umbilical cord opening and allowed to flow or percolate out of openings in the wall of the placenta which interfaced with the maternal uterine wall. In another embodiment, the perfusion solution is passed through the umbilical veins and collected from the umbilical artery, or is passed through the umbilical artery and collected from the umbilical veins, that is, is passed through only the placental vasculature (fetal tissue).
[0074] In one embodiment, for example, the umbilical artery and the umbilical vein are connected simultaneously, e.g., to a pipette that is connected via a flexible connector to a reservoir of the perfusion solution. The perfusion solution is passed into the umbilical vein and artery. The perfusion solution exudes from and/or passes through the walls of the blood vessels into the surrounding tissues of the placenta, and is collected in a suitable open vessel from the surface of the placenta that was attached to the uterus of the mother during gestation. The perfusion solution may also be introduced through the umbilical cord opening and allowed to flow or percolate out of openings in the wall of the placenta which interfaced with the maternal uterine wall. Placental cells that are collected by this method, which can be referred to as a "pan" method, are typically a mixture of fetal and maternal cells.
[0075] In another embodiment, the perfusion solution is passed through the umbilical veins and collected from the umbilical artery, or is passed through the umbilical artery and collected from the umbilical veins. Placental cells collected by this method, which can be referred to as a "closed circuit" method, are typically almost exclusively fetal.
[0076] The closed circuit perfusion method can, in one embodiment, be performed as follows. A post-partum placenta is obtained within about 48 hours after birth. The umbilical cord is clamped and cut above the clamp. The umbilical cord can be discarded, or can processed to recover, e.g., umbilical cord stem cells, and/or to process the umbilical cord membrane for the production of a biomaterial. The amniotic membrane can be retained during perfusion, or can be separated from the chorion, e.g., using blunt dissection with the fingers. If the amniotic membrane is separated from the chorion prior to perfusion, it can be, e.g., discarded, or processed, e.g., to obtain stem cells by enzymatic digestion, or to produce, e.g., an amniotic membrane biomaterial, e.g., the biomaterial described in U.S. Application Publication No.
2004/0048796, the disclosure of which is hereby incorporated by reference herein in its entirety. After cleaning the placenta of all visible blood clots and residual blood, e.g., using sterile gauze, the umbilical cord vessels are exposed, e.g., by partially cutting the umbilical cord membrane to expose a cross-section of the cord. The vessels are identified, and opened, e.g., by advancing a closed alligator clamp through the cut end of each vessel. The apparatus, e.g., plastic tubing connected to a perfusion device or peristaltic pump, is then inserted into each of the placental arteries. The pump can be any pump suitable for the purpose, e.g., a peristaltic pump. Plastic tubing, connected to a sterile collection reservoir, e.g., a blood bag such as a 250 mL collection bag, is then inserted into the placental vein. Alternatively, the tubing connected to the pump is inserted into the placental vein, and tubes to a collection reservoir(s) are inserted into one or both of the placental arteries. The placenta is then perfused with a volume of perfusion solution, e.g., about 750 ml of perfusion solution. Cells in the perfusate are then collected, e.g., by
centrifugation.
[0077] In one embodiment, the proximal umbilical cord is clamped during perfusion, and, more specifically, can be clamped within 4-5 cm (centimeter) of the cord' s insertion into the placental disc.
[0078] The first collection of perfusion fluid from a mammalian placenta during the exsanguination process is generally colored with residual red blood cells of the cord blood and/or placental blood. The perfusion fluid becomes more colorless as perfusion proceeds and the residual cord blood cells are washed out of the placenta. Generally from 30 to 100 mL of perfusion fluid is adequate to initially flush blood from the placenta, but more or less perfusion fluid may be used depending on the observed results.
[0079] In certain embodiments, cord blood is removed from the placenta prior to perfusion (e.g., by gravity drainage), but the placenta is not flushed (e.g., perfused) with solution to remove residual blood. In certain embodiments, cord blood is removed from the placenta prior to perfusion (e.g., by gravity drainage), and the placenta is flushed (e.g., perfused) with solution to remove residual blood.
[0080] The volume of perfusion liquid used to perfuse the placenta may vary depending upon the number of placental cells to be collected, the size of the placenta, the number of collections to be made from a single placenta, etc. In various embodiments, the volume of perfusion liquid may be from 50 mL to 5000 mL, 50 mL to 4000 mL, 50 mL to 3000 mL, 100 mL to 2000 mL, 250 mL to 2000 mL, 500 mL to 2000 mL, or 750 mL to 2000 mL. Typically, the placenta is perfused with 700-800 mL of perfusion liquid following exsanguination.
[0081] The placenta can be perfused a plurality of times over the course of several hours or several days. Where the placenta is to be perfused a plurality of times, it may be maintained or cultured under aseptic conditions in a container or other suitable vessel, and perfused with a cell collection composition, or a standard perfusion solution (e.g., a normal saline solution such as phosphate buffered saline ("PBS") with or without an anticoagulant (e.g., heparin, warfarin sodium, coumarin, bishydroxycoumarin), and/or with or without an antimicrobial agent (e.g., β- mercaptoethanol (0.1 mM); antibiotics such as streptomycin (e.g., at 40-100 μg/ml), penicillin (e.g., at 40 U/ml), amphotericin B (e.g., at 0.5 μg/ml). In one embodiment, an isolated placenta is maintained or cultured for a period of time without collecting the perfusate, such that the placenta is maintained or cultured for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 hours, or 2 or 3 or more days before perfusion and collection of perfusate. The perfused placenta can be maintained for one or more additional time(s), e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or more hours, and perfused a second time with, e.g., 700-800 mL perfusion fluid. The placenta can be perfused 1, 2, 3, 4, 5 or more times, for example, once every 1, 2, 3, 4, 5 or 6 hours. In one embodiment, perfusion of the placenta and collection of perfusion solution, e.g., placental cell collection composition, is repeated until the number of recovered nucleated cells falls below 100 cells/ml. The perfusates at different time points can be further processed individually to recover time-dependent populations of cells, e.g., total nucleated cells. Perfusates from different time points can also be pooled.
4.3. Isolated Placental Cells
[0082] The placental tissue banked according to the methods described herein may be one or more isolated placental cells. Said isolated placental cells may be derived from the placenta according to any of the methods described herein (e.g., by perfusion).
[0083] The isolated placental cells banked in accordance with the methods described herein may be hematopoietic cells, natural killer cells isolated from placental perfusate, placental intermediate natural killer (Pi K) cells, placenta-derived adherent cells, or amnion-derived adherent cells.
4.3.1.1. Hematopoietic Stem Cells
[0084] The isolated placental cells banked in accordance with the methods described herein may comprise hematopoietic stem cells. In one embodiment, said hematopoietic stem cells are isolated from placental perfusate. In another embodiment, said hematopoietic stem cells are CD34+. In another embodiment, said CD34+ hematopoietic stem cells comprise at least 50% of said isolated placental stem cells.
4.3.1.2. Placenta-Derived Adherent Cells
[0085] The isolated placental cells banked in accordance with the methods described herein may be placenta-derived adherent cells (e.g., the placenta-derived adherent cells disclosed in U.S. Patent Nos. 7,468,276; 8,057,788 and 8,202,703, the disclosures of which are hereby
incorporated by reference in their entireties). [0086] In a specific embodiment, the placenta-derived adherent cells are CD34", CD10+, CD105+ and CD200+ tissue culture plastic placental adherent cells.
[0087] In another specific embodiment, the placenta-derived adherent cells express one or more genes at a level at least two-fold higher than bone marrow-derived mesenchymal stem cells, wherein said genes are selected from the group consisting of ACTG2, ADARB1, AMIG02, ARTS-1, B4GALT6, BCHE, Cl lorf9, CD200, COL4A1, COL4A2, CPA4, DMD, DSC3, DSG2, ELOVL2, F2RL1, FIJ10781, GATA6, GPR126, GPRC5B, HLA-G, ICAM1, IER3, IGFBP7, ILIA, IL6, IL18, KRT18, KRT8, LIPG, LRAP, MATN2, MEST, FE2L3, NUAK1, PCDH7, PDLIM3, PJP2, RTN1, SERPINB9, ST3GAL6, ST6GALNAC5, SLC12A8, TCF21, TGFB2, VTN, and ZC3H12A. In another embodiment, the placenta-derived adherent cells express one or more genes at a level at least two-fold higher than bone marrow-derived mesenchymal stem cells, wherein said one or more genes are one or more of LOVL2, ST3GAL6, ST6GALNAC5, and/or SLC12A8. In another embodiment, the placenta-derived adherent cells express one or more genes at a level at least two-fold higher than bone marrow-derived mesenchymal stem cells, wherein said one or more genes are one or more of CPA4, TCF21, VTN, B4GALT6, FLJ 10781, and/or NUAK1.
[0088] In certain embodiments, the placenta-derived adherent cells are grown in culture, e.g., the placenta-derived adherent cells have adhered to a tissue culture substrate. In a specific embodiment, the cells are cultured in the presence of serum. In another specific embodiment, the cells are cultured in the absence of serum.
[0089] In one embodiment, the placenta-derived adherent cells are substantially fetal in origin, e.g., the placenta-derived adherent cells are about or at least 90%, 95%, 99% or 100% fetal in origin. In another embodiment, the placenta-derived adherent cells are of mixed origin, e.g., the placenta-derived adherent cells are both fetal and maternal in origin.
[0090] In certain embodiments, the placenta-derived adherent cells have been passaged prior to isolation of exosomes from the placenta-derived adherent cells. In a specific embodiment, the placenta-derived adherent cells have been passaged about or at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, or 20 times. In a specific embodiment, the placenta-derived adherent cells have been cultured in vitro for 6 passages.
[0091] In certain embodiments, the placenta-derived adherent cells have been expanded in vitro. In a specific embodiment, the placenta-derived adherent cells have been expanded for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38 or 40 population doublings, or more.
4.3.1.3. Natural Killer Cells
[0092] The isolated placental cells banked in accordance with the methods described herein may be natural killer (NK) cells. In a specific embodiment, said NK cells are isolated from placental perfusate.
4.3.1.3.1. Placental Intermediate Natural Killer (PiNK) Cells
[0093] In some embodiments, the natural killer cells banked in accordance with the methods described herein are placental intermediate natural killer (PiNK) cells (see also U.S. Patent No. 8,263,065, the disclosure of which is hereby incorporated by reference in its entirety). In various embodiments, PiNK cells are derived from placental cells. In specific embodiments, the placental cells are obtained from placental perfusate, e.g., human placental perfusate. In specific embodiments, the placental cells are obtained from placental tissue that has been mechanically and/or enzymatically disrupted.
[0094] PiNK cells are characterized as being CD56+ CD16", i.e., displaying the CD56 cellular marker and lacking the CD 16 cellular marker, e.g., as determined by flow cytometry, e.g., fluorescence-activated cell sorting using antibodies against CD 16 and CD56, as described above.
[0095] In certain embodiments, the PiNK cells are CD3~.
[0096] In other embodiments, the PiNK cells do not exhibit one or more cellular markers exhibited by fully mature natural killer cells (e.g., CD 16), or exhibit such one or more markers at a detectably reduced level compared to fully mature natural killer cells, or exhibit one or more cellular markers associated with natural killer cell precursors but not fully mature natural killer cells. In a specific embodiment, a PiNK cell described herein expresses NKG2D, CD94 and/or NKp46 at a detectably lower level than a fully mature NK cell. In another specific embodiment, a plurality of PiNK cells described herein expresses, in total, NKG2D, CD94 and/or NKp46 at a detectably lower level than an equivalent number of fully mature NK cells.
[0097] In certain embodiments, PiNK cells express one or more of the microRNAs hsa-miR- 100, hsa-miR-127, hsa-miR-211, hsa-miR-302c, hsa-miR-326, hsa-miR-337, hsa-miR-497, hsa- miR-512-3p, hsa-miR-515-5p, hsa-miR-517b, hsa-miR-517c, hsa-miR-518a, hsa-miR-518e, hsa- miR-519d, hsa-miR-520g, hsa-miR-520h, hsa-miR-564, hsa-miR-566, hsa-miR-618, and/or hsa- miR-99a at a detectably higher level than peripheral blood natural killer cells. [0098] Because the post-partum placenta comprises tissue and cells from the fetus and from the maternal placental perfusate, depending upon the method of collection, Pi K cells can comprise fetal cells only, or a substantial majority of fetal cells (e.g., greater than about 90%, 95%, 98%> or 99%), or can comprise a mixture of fetal and maternal cells (e.g., the fetal cells comprise less than about 90%, 80%, 70%, 60%, or 50% of the total nucleated cells of the perfusate). In one embodiment, the PiNK cells are derived only from fetal placental cells, e.g., cells obtained from closed-circuit perfusion of the placenta (see above) wherein the perfusion produces perfusate comprising a substantial majority, or only, fetal placental cells. In another embodiment, the PiNK cells are derived from fetal and maternal cells, e.g., cells obtained by perfusion by the pan method (see above), wherein the perfusion produced perfusate comprising a mix of fetal and maternal placental cells. Thus, in one embodiment, the NK cells are a population of placenta- derived intermediate natural killer cells, the substantial majority of which have the fetal genotype. In another embodiment, the NK cells are a population of placenta-derived intermediate natural killer cells that comprise natural killer cells having the fetal genotype and natural killer cells having the maternal phenotype.
4.3.1.4. Amnion Derived Adherent Cells
[0099] The isolated placental stem cells banked in accordance with the methods described herein can comprise amnion derived adherent cells (AMDACs), e.g., the amnion derived cells described in U.S. Patent No. 8,367,409, the disclosure of which is hereby incorporated by reference in its entirety.
[00100] Amnion derived adherent cells, and isolated populations of cells comprising the amnion derived adherent cells, provided herein can be produced by, e.g., digestion of amnion tissue followed by selection for adherent cells. In one embodiment, for instance, isolated amnion derived adherent cells, or an isolated population of cells comprising amnion derived adherent cells, can be produced by (1) digesting amnion tissue with a first enzyme to dissociate cells from the epithelial layer of the amnion from cells from the mesenchymal layer of the amnion; (2) subsequently digesting the mesenchymal layer of the amnion with a second enzyme to form a single-cell suspension; (3) culturing cells in said single-cell suspension on a tissue culture surface, e.g., tissue culture plastic; and (4) selecting cells that adhere to said surface after a change of medium, thereby producing an isolated population of cells comprising amnion derived adherent cells. In a specific embodiment, said first enzyme is trypsin. In a more specific embodiment, said trypsin is used at a concentration of 0.25% trypsin (w/v), in 5-20, e.g., 10 milliliters solution per gram of amnion tissue to be digested. In another more specific
embodiment, said digesting with trypsin is allowed to proceed for about 15 minutes at 37°C and is repeated up to three times. In another specific embodiment, said second enzyme is
collagenase. In a more specific embodiment, said collagenase is used at a concentration between 50 and 500 U/L in 5 mL per gram of amnion tissue to be digested. In another more specific embodiment, said digesting with collagenase is allowed to proceed for about 45-60 minutes at 37°C. In another specific embodiment, the single-cell suspension formed after digestion with collagenase is filtered through, e.g., a 75 μΜ - 150 μΜ filter between step (2) and step (3). In another specific embodiment, said first enzyme is trypsin, and said second enzyme is collagenase.
[00101] Amnion derived adherent cells superficially resemble mesenchymal cells in
appearance, having a generally fibroblastoid shape. The cells adhere to a cell culture surface, e.g., to tissue culture plastic.
[00102] AMDACs display cellular markers that distinguish them from other amnion-derived, or placenta-derived, cells. For example, in one embodiment, the amnion derived adherent cell is OCT-4" (octamer binding protein 4), as determined by RT-PCR. In another specific embodiment, the OCT-4" amnion derived adherent cell is O49 , as determined by immunolocalization. In another specific embodiment, said OCT-4" cell is HLA-G", as determined by RT-PCR. In another specific embodiment, the OCT-4" cell is VEGFR1/Flt-1+ (vascular endothelial growth factor receptor 1) and/or VEGFR2/KDR+ (vascular endothelial growth factor receptor 2), as determined by immunolocalization. In a specific embodiment, the OCT-4" amnion derived adherent cell, or a population of OCT-4" amnion derived adherent cells, expresses at least 2 log less PCR-amplified mRNA for OCT-4 at, e.g., 20 cycles, than an NTERA-2 cell, or population of NTERA-2 cells having an equivalent number of cells and RNA amplification cycles. In another specific embodiment, said OCT-4" cell is CD90+, CD105+, or CD117". In a more specific embodiment, said OCT-4" cell is CD90+, CD105+, and CD117". In a more specific embodiment, the cell is OCT-4" or HLA-G-, and is additionally CD49 , CD90+, CD105+, and CD117". In a more specific embodiment, the cell is OCT-4", HLA-G", CD49f*, CD90+, CD105+, and CD117". In another specific embodiment, the OCT-4" cell does not express SOX2, e.g., as determined by RT-PCR for 30 cycles. In a specific embodiment, therefore, the cell is OCT-4", O49 , CD90+, CD105+, and CD117", as determined by immunolocalization or flow cytometry, and SOX2 , as determined by RT-PCR, e.g., for 30 cycles. In a specific embodiment, the cell is OCT-4" as determined by RT-PCR, CD49 , CD90+, and HLA-G" as determined by flow cytometry.
[00103] In another embodiment, said OCT-4" cell is one or more of CD29+, CD73+, ABC-p+, and CD38", as determined by immunolocalization.
5. Examples
5.1. Example 1: Method of Banking Placental Material
[00104] This example provides a method for banking placental tissue and the creation of a bank for subsequently retrieving placental tissue and one or more cell types or related tissues.
5.1.1. Processing of Placental Tissue
5.1.1.1. Whole Placenta
[00105] An intact placenta to be stored in a placental tissue bank is received by the bank directly from the customer, or from a source that is not the customer. Placenta donors are recruited from expectant mothers that are enrolled in private umbilical cord blood banking programs and provide informed consent permitting the use of the exsanguinated placenta following recovery of cord blood for research purposes. Donor data may be confidential. The donor may be a customer of the bank, or the placenta may be donated to the bank by an individual who is not a customer of the bank. These donors also permit use of blinded data generated from the normal processing of their umbilical cord blood specimens for
cryopreservation. This allows comparison between the composition of the collected cord blood and the effluent perfusate recovered using the experimental method described below.
[00106] Following exsanguination of cord blood from the umbilical cord and placenta, the placenta is stored at room temperature and delivered to the laboratory within four to twenty-four hours. Alternatively, the placenta is placed in a sterile, insulated container at room temperature and delivered to the laboratory within 4 hours of birth. Placentas are discarded if, on inspection, they have evidence of physical damage such as fragmentation of the organ or avulsion of umbilical vessels. Placentas are maintained at room temperature (23±2°C) or refrigerated (4°C) in sterile containers for 2 to 20 hours. Periodically, the placentas are immersed and washed in sterile saline at 25±3 °C to remove any visible surface blood or debris. [00107] The umbilical cord is transected approximately 5 cm from its insertion into the placenta and the umbilical vessels are cannulated with TEFLON® or polypropylene catheters connected to a sterile fluid path allowing bi-directional perfusion of the placenta and recovery of the effluent fluid. Accordingly, the various aspects of placental conditioning, perfusion and effluent collection to be performed under controlled ambient atmospheric conditions are monitored, as is the real-time monitoring of intravascular pressure and flow rates, core and perfusate temperatures and recovered effluent volumes. A range of conditioning protocols are evaluated over a 24-hour postpartum period, and the cellular composition of the effluent fluid is analyzed by flow cytometry, light microscopy and colony forming unit assays.
5.1.1.2. Placental Conditioning
[00108] The donor placentas are processed at room temperature within 12 to 24 hours after delivery. Before processing, the membranes are removed and the maternal site washed clean of residual blood. The umbilical vessels are cannulated with catheters made from 20 gauge Butterfly needles use for blood sample collection.
[00109] The donor placentas are maintained under varying conditions such as maintenance at 5-37° 5% C02, pH 7.2 to 7.5, in an attempt to simulate and sustain a physiologically compatible environment for the proliferation and recruitment of residual embryonic-like stem cells, if necessary. The cannula is flushed with IMDM serum-free medium (GibcoBRL, NY) containing 2U/ml heparin (Elkins-Sinn, NJ). Perfusion of the placenta continues at a rate of 50 ml per minute until approximately 150 ml of perfusate is collected. This volume of perfusate is labeled "early fraction." Continued perfusion of the placenta at the same rate results in the collection of a second fraction of approximately 150 ml and is labeled "late fraction." During the course of the procedure, the placenta is gently massaged to aid in the perfusion process and assist in the recovery of cellular material. Effluent fluid is collected from the perfusion circuit by both gravity drainage and aspiration through the arterial cannula.
5.1.1.3. Placental Perfusate Cell Isolation
[00110] This example describes the method of recovering placental perfusate and total nucleated cells from placental perfusate, and banking the isolated placental tissue.
[00111] Twenty ml of phosphate buffered saline solution (PBS) are added to the perfusion liquid and a 10 ml portion is collected and centrifuged for 25 minutes at 3000 rpm (revolutions per minute). The effluent is divided into four tubes and placed in an ice bath. 2.5 ml of a 1% fetal calf serum (FCS) solution in PBS are added and the tubes are centrifuged (140 minutes x 10 g (acceleration due to gravity)). The pellet is resuspended in 5 ml of 1% FCS and two tubes are combined. The total mononucleocytes are calculated by adding the total lymphocytes and the total monocytes, and then multiplying the result by the total cell suspension volume.
5.1.1.4. Additional Placental and Embryonic Tissue
[00112] In addition to the placental perfusate and/or total nucleated cells isolated from a perfused placenta, distinct populations of fetal and/or maternal tissue are obtained from the placentas and umbilical cords of normal, full-term pregnancies. All donors provide full written consent for the use of their placentas for research purposes. Isolation of the following tissues is accomplished by dissection: (1) whole umbilical cord, (2) amniotic membrane, (3) chorionic membrane, (4) umbilical cord vessels, (5) umbilical cord membrane, and (6) placental vessels. Umbilical cord blood and platelet-rich plasma from placental blood or umbilical cord blood are isolated by collecting the blood directly from the placenta or umbilical cord using standard techniques known in the art.
5.1.1.5. Isolated Placental Stem Cells
[00113] In addition to cells isolated from placental perfusate as described in Section 5.1.1.2, Supra, placental stem cells are obtained from the following sources: enzymatic digestions of (1) amnion, (2) chorion, (3) amnion-chorion plate, and (4) umbilical cord. The various placental tissue are cleaned in sterile PBS (Gibco-Invitrogen Corporation, Carlsbad, CA) and placed on separate sterile Petri dishes. The various tissues are minced using a sterile surgical scalpel and placed into 50 mL Falcon Conical tubes. The minced tissues are digested with IX Collagenase (Sigma- Aldrich, St. Louis, MO) for 20 minutes in a 37°C water bath, centrifuged, and then digested with 0.25% Trypsin-EDTA (Gibco-Invitrogen Corp) for 10 minutes in a 37°C water bath. The various tissues are centrifuged after digestion and rinsed once with sterile PBS (Gibco-Invitrogen Corp). The reconstituted cells are then filtered twice, once with 100 μπι cell strainers and once with 30 μπι separation filters, to remove any residual extracellular matrix or cellular debris. The isolated cells are cultured according to the methods described herein and cultured as appropriate to yield the one or more isolated placental stem cells (See Section 4.4, Supra). 5.1.2. Cryopreservation of Placenta and Isolated Cells
[00114] Placental tissue and one or more additional tissues derived from the placenta and/or umbilical cord are isolated and prepared according to the methods in Section 5.1.1.
[00115] Method of cryopreserving tissues and cells are well known in the art. In one embodiment, the whole placenta and other placental tissue are stored at -80°C to -86°C for short periods of time, e.g., for 10 minutes, 30 minutes, 60 minutes, two hours, three hours, four hours, or more. The whole placenta and/or other placental tissue are then transferred to liquid nitrogen for permanent storage. Prior to the cryopreservation of a whole placenta, the whole placenta may be perfused, e.g., perfused with a saline solution comprising dimethyl sulfoxide (DMSO).
[00116] Cells to be cryopreserved are harvested from culture with Trypsin-EDTA, quenched with 2% FBS in PBS, and counted on a hemacytometer. After centrifugation, cells are resuspended with 10% DMSO in FBS to a concentration of about 1 million cells/ml for cells to be used for assembly of a cell bank, and 10 million cells/ml for individual frozen cell doses. The cell solution is transferred to a freezing container, which is placed in an isopropyl alcohol bath in a -80°C freezer. The following day, cells are transferred to liquid nitrogen.
5.1.3. Identifier Assignment for Creation of a Placental Tissue Bank
[00117] The intact placenta and each additional cell or tissue type is given a unique identifier that is stored by the bank prior to the banking of the cell or tissue itself. The identifier is specific to a single sample and an individual customer, and after assignment the identifier is conveyed to the customer and stored by the bank. The stored identifier(s) allows for future retrieval of each one or more sample from the bank at the request of the customer. In the event a customer elects to bank more than one sample (e.g., an intact placenta and additionally a population of isolated placental stem cells), the first sample receives a first identifier, and the second sample receives a second identifier. The customer may elect to bank the one or more samples prior to the bank receiving the placental tissue or prior to the processing of the placental tissue. In the event that a customer elects a cell or tissue type prior to the receipt of the placental tissue or processing of placental tissue, the elected tissue or cell type is assigned a third identifier. The placental tissue not elected by the customer are then assigned a fourth identifier to specify non-election. Non- elected cells and tissues may then be discarded or further stored by the bank for use by future customers or other purposes. [00118] The assignment of identifiers by the customer allows for the specific banking of placental tissue according to the customer's preferences. It also permits customer-specific retrieval of the banked placental tissue at the customer's request.
Equivalents:
[00119] The present disclosure is not to be limited in scope by the specific embodiments described herein. Indeed, various modifications of the subject matter provided herein, in addition to those described, will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications are intended to fall within the scope of the appended claims.
[00120] Various publications, patents and patent applications are cited herein, the disclosures of which are incorporated by reference in their entireties.

Claims

What is Claimed:
1. A method of banking placental tissue in a placental tissue bank, comprising:
a. receiving a placenta or a portion thereof;
b. processing the placenta to isolate the placental tissue; and
c. banking the placental tissue.
2. The method of claim 1, wherein the placental tissue comprises whole placenta.
3. The method of claim 1, wherein the placental tissue comprises placental perfusate or total nucleated cells isolated from placental perfusate.
4. The method of claim 3, wherein the placental tissue additionally comprises one or more of:
a. whole umbilical cord;
b. amniotic membrane;
c. chorionic membrane;
d. umbilical cord vessels;
e. umbilical cord membrane;
f. placental vessels;
g. platelet-rich plasma from placental blood or umbilical cord blood;
h. isolated placental cells.
5. The method of any of claims 1-4, wherein a customer of said placental tissue bank directs which of said tissues are banked by the placental tissue bank.
6. The method of any of claims 1-4, wherein said tissue is received from a customer of the placental tissue bank.
7. The method of any of claims 1-4, wherein said tissue is received from a source other than a customer of the placental tissue bank.
8. The method of claim 3, wherein said placental perfusate has been collected from a placenta that has been exsanguinated.
9. The method of claim 3, wherein said placental perfusate has been collected from a placenta that has been exsanguinated but not perfused to remove residual blood.
10. The method of claim 3, wherein said placental perfusate has been collected from a placenta that has not been exsanguinated.
11. The method of claim 4, wherein said isolated placental cells comprise CD34+
hematopoietic stem cells isolated from placental perfusate, wherein the CD34+ hematopoietic stem cells comprise at least 50% of said isolated placental cells.
12. The method of claim 4, wherein said isolated placental cells comprise natural killer cells isolated from placental perfusate, wherein the natural killer cells comprise at least 50% of said isolated placental cells.
13. The method of claim 12, wherein said natural killer cells are placental intermediate natural killer (Pi K) cells.
14. The method of claim 4, wherein said isolated placental cells are CD 10 , CD34", CD 105 , CD200+ tissue culture plastic placental adherent cells.
15. The method of claim 4, wherein said isolated placental cells are amni on-derived adherent cells.
16. The method of claim 3, wherein said placental perfusate is combined with umbilical cord blood.
17. The method of claim 3, wherein said total nucleated cells from placental perfusate are combined with umbilical cord blood.
18. The method of claim 3, wherein said placental perfusate is combined with total nucleated cells from umbilical cord blood.
19. The method of claim 3, wherein said total nucleated cells from placental perfusate are combined with total nucleated cells from umbilical cord blood.
20. The method of claim 2, wherein said whole placenta is decellularized.
21. The method of claim 1 or claim 4, wherein said placental tissue is amniotic membrane.
22. The method of claim 21, wherein said amniotic membrane is decellularized.
23. The method of claim 1 or claim 4, wherein said placental tissue is chorionic membrane.
24. The method of claim 23, wherein said chorionic membrane is decellularized.
25. The method of claim 1 or claim 4, wherein said placental tissue is umbilical cord.
26. The method of claim 25, wherein said umbilical cord is decellularized.
27. The method of claim 1 or claim 4, wherein said placental tissue is umbilical cord vessels or placental vessels.
28. The method of claim 27, wherein said vessels are decellularized.
29. The method of claim 1 or claim 4, wherein said placental tissue is umbilical cord membrane.
30. The method of claim 29, wherein said umbilical cord membrane is decellularized.
31. The method of any of claims 1-30, wherein said banking of placental tissue is adjunct to banking of placental blood and/or umbilical cord blood.
32. The method of claim 1 or claim 3, wherein the method comprises banking of (i) at least one unit of placental blood or umbilical cord blood and (ii) at least one unit of perfusate or HPDSC, wherein said at least one unit of placental blood or umbilical cord blood, and said at least one unit of perfusate or HPDSC are from the same placenta.
33. The method of any of claims 1-30, wherein said placental tissue is banked without cryopreservation.
34. The method of any of claims 1-30, wherein said placental tissue is cryopreserved for banking.
35. The method of claim 34, wherein said placental tissue is cryopreserved by the placental tissue bank.
36. The method of any of claims 1-30, wherein said placental tissue is allogeneic to a customer of said placental tissue bank.
37. The method of any of claims 1-30, wherein said placental tissue is autologous to a customer of said placental tissue bank.
38. The method of any of claims 1-37, comprising assigning the placenta and/or placental tissue an identifier based on the source of the placenta.
39. The method of claim 38, comprising assigning the placenta a first identifier, and assigning each of said placental tissue a second tissue-specific identifier.
40. The method of claim 38, wherein said second tissue-specific identifier incorporates said first identifier.
41. The method of claim 39 or claim 40, wherein said placental tissue bank comprises a system for storing and retrieving said first and second identifiers.
42. The method of any of claims 1-4 wherein the method comprises receiving, prior to step (a) or step (b), an election from the customer as to which placental tissue from said placenta to bank.
43. The method of claim 42, wherein the placental tissue elected by the customer are assigned a third identifier to indicate election.
44. The method of claim 44, wherein placental tissue not elected by the customer are assigned a fourth identifier to indicate non-election.
45. A method of providing placental tissue to a customer, comprising:
a. providing the customer a selection of placental tissue from said placenta or portion thereof to bank, wherein the tissues comprise (1) one or both of whole placenta placental perfusate, and/or cells isolated from placental perfusate, and (2) additionally one or more of:
i. amniotic membrane;
ii. chorionic membrane;
iii. whole umbilical cord;
iv. umbilical cord vessels;
v. umbilical cord membrane;
vi. placental vessels;
vii. platelet-rich plasma from placental blood or umbilical cord blood;
viii. isolated placental cells;
b. receiving an election from the customer as to which placental tissue to bank; c. processing the placenta to isolate the elected tissue(s); and
d. banking the tissues at the direction of said customer, thereby providing placental tissue to a customer.
46. The method of claim 45, wherein the placental tissue comprises whole placenta.
47. The method of claim 45, wherein the placental tissue comprises placental perfusate or total nucleated cells isolated from placental perfusate.
48. The method of claim 47, wherein the placental tissue additionally comprises one or more of:
a. whole umbilical cord;
b. amniotic membrane;
c. chorionic membrane;
d. umbilical cord vessels;
e. umbilical cord membrane;
f. placental vessels;
g. platelet-rich plasma from placental blood or umbilical cord blood;
h. isolated placental cells.
49. The method of any of claims 45-48, wherein a customer of said placental tissue bank directs which of said tissues are banked by the placental tissue bank.
50. The method of any of claims 45-48, wherein said tissue is received from a customer of the placental tissue bank.
51. The method of any of claims 45-48, wherein said tissue is received from a source other than a customer of the placental tissue bank.
52. The method of claim 47, wherein said placental perfusate has been collected from a placenta that has been exsanguinated.
53. The method of claim 47, wherein said placental perfusate has been collected from a placenta that has been exsanguinated but not perfused to remove residual blood.
54. The method of claim 47, wherein said placental perfusate has been collected from a placenta that has not been exsanguinated.
55. The method of claim 48, wherein said isolated placental cells comprise CD34+
hematopoietic stem cells isolated from placental perfusate, wherein the CD34+ hematopoietic stem cells comprise at least 50% of said isolated placental cells.
56. The method of claim 48, wherein said isolated placental cells comprise natural killer cells isolated from placental perfusate, wherein the natural killer cells comprise at least 50% of said isolated placental cells.
57. The method of claim 56, wherein said natural killer cells are placental intermediate natural killer (Pi K) cells.
58. The method of claim 48, wherein said isolated placental cells are CD 10 , CD34", CD 105 , CD200+ tissue culture plastic placental adherent cells.
59. The method of claim 48, wherein said isolated placental cells are amnion-derived adherent cells.
60. The method of claim 47, wherein said placental perfusate is combined with umbilical cord blood.
61. The method of claim 47, wherein said total nucleated cells from placental perfusate are combined with umbilical cord blood.
62. The method of claim 47, wherein said placental perfusate is combined with total nucleated cells from umbilical cord blood.
63. The method of claim 47, wherein said total nucleated cells from placental perfusate are combined with total nucleated cells from umbilical cord blood.
64. The method of claim 46, wherein said whole placenta is decellularized.
65. The method of claim 45 or claim 48, wherein said placental tissue is amniotic membrane.
66. The method of claim 65, wherein said amniotic membrane is decellularized.
67. The method of claim 45 or claim 48, wherein said placental tissue is chorionic membrane.
68. The method of claim 67, wherein said chorionic membrane is decellularized.
69. The method of claim 45 or claim 48, wherein said placental tissue is umbilical cord.
70. The method of claim 69, wherein said umbilical cord is decellularized.
71. The method of claim 45 or claim 48, wherein said placental tissue is umbilical cord vessels or placental vessels.
72. The method of claim 71, wherein said vessels are decellularized.
73. The method of claim 45 or claim 48, wherein said placental tissue is umbilical cord membrane.
74. The method of claim 73, wherein said umbilical cord membrane is decellularized.
75. The method of any of claims 45-74, wherein said banking of placental tissue is adjunct to banking of placental blood and/or umbilical cord blood.
76. The method of claim 45 or claim 48, wherein the method comprises banking of (i) at least one unit of placental blood or umbilical cord blood and (ii) at least one unit of perfusate or HPDSC, wherein said at least one unit of placental blood or umbilical cord blood, and said at least one unit of perfusate or HPDSC are from the same placenta.
77. The method of any of claims 45-74, wherein said placental tissue is banked without cryopreservation.
78. The method of any of claims 45-74, wherein said placental tissue is cryopreserved for banking.
79. The method of claim 78, wherein said placental tissue is cryopreserved by the placental tissue bank.
80. The method of any of claims 45-74, wherein said placental tissue is allogeneic to a customer of said placental tissue bank.
81. The method of any of claims 45-74, wherein said placental tissue is autologous to a customer of said placental tissue bank.
82. The method of any of claims 45-81, comprising assigning the placenta and/or placental tissue an identifier based on the source of the placenta.
83. The method of claim 82, comprising assigning the placenta a first identifier, and assigning each of said placental tissue a second tissue-specific identifier.
84. The method of claim 82, wherein said second tissue-specific identifier incorporates said first identifier.
85. The method of claim 83 or claim 84, wherein said placental tissue bank comprises a system for storing and retrieving said first and second identifiers.
86. The method of any of claims 45-48 wherein the method comprises receiving, prior to step (a) or step (b), an election from the customer as to which placental tissue from said placenta to bank.
87. The method of claim 86, wherein the placental tissue elected by the customer are assigned a third identifier to indicate election.
88. The method of claim 86, wherein placental tissue not elected by the customer are assigned a fourth identifier to indicate non-election.
89. The method of claim 45, additionally comprising providing umbilical cord blood or placental blood to said customer.
A method of banking placental tissue in a placental tissue bank, comprising:
a. receiving a placenta or a portion thereof from a customer;
b. obtaining placental blood and/or umbilical cord from said placenta;
c. providing a customer with a selection of placental tissue from said placenta or portion thereof to bank;
d. receiving an election from the customer as to which placental tissue to bank; e. processing the placenta to isolate the elected tissue(s); and
f. banking the tissues, wherein the tissues comprise one or more of:
i. whole placenta;
ii. amniotic membrane;
iii. chorionic membrane;
iv. whole umbilical cord;
v. umbilical cord vessels;
vi. umbilical cord membrane;
vii. placental vessels;
viii. platelet-rich plasma from placental blood or umbilical cord blood;
ix. isolated placental cells;
x. placental perfusate;
xi. human placenta-derived stem cells; or
xii. cells isolated from placental perfusate; and
g. optionally banking said umbilical cord blood or said placental blood.
The method of claim 90, wherein if the customer selects the tissue for banking to be one more of whole placenta, amniotic membrane, chorionic membrane, whole umbilical cord, umbilical cord vessels, umbilical cord membrane, placental vessels, additionally comprising providing the customer with the option of decellularizing the tissue.
92. The method of claim 91, wherein the placental tissue comprises whole placenta.
93. The method of claim 91, wherein the placental tissue comprises placental perfusate or total nucleated cells isolated from placental perfusate.
94. The method of claim 93, wherein the placental tissue additionally comprises one or more of:
a. whole umbilical cord;
b. amniotic membrane;
c. chorionic membrane;
d. umbilical cord vessels;
e. umbilical cord membrane;
f. placental vessels;
g. platelet-rich plasma from placental blood or umbilical cord blood;
h. isolated placental cells.
95. The method of any of claims 91-94, wherein a customer of said placental tissue bank directs which of said tissues are banked by the placental tissue bank.
96. The method of any of claims 91-94, wherein said tissue is received from a customer of the placental tissue bank.
97. The method of any of claims 91-94, wherein said tissue is received from a source other than a customer of the placental tissue bank.
98. The method of claim 93, wherein said placental perfusate has been collected from a placenta that has been exsanguinated.
99. The method of claim 93, wherein said placental perfusate has been collected from a placenta that has been exsanguinated but not perfused to remove residual blood.
100. The method of claim 93, wherein said placental perfusate has been collected from a placenta that has not been exsanguinated.
101. The method of claim 94, wherein said isolated placental cells comprise CD34+
hematopoietic stem cells isolated from placental perfusate, wherein the CD34+ hematopoietic stem cells comprise at least 50% of said isolated placental cells.
102. The method of claim 94, wherein said isolated placental cells comprise natural killer cells isolated from placental perfusate, wherein the natural killer cells comprise at least 50% of said isolated placental cells.
103. The method of claim 102, wherein said natural killer cells are placental intermediate natural killer (Pi K) cells.
104. The method of claim 94, wherein said isolated placental cells are CD 10 , CD34", CD 105 , CD200+ tissue culture plastic placental adherent cells.
105. The method of claim 94, wherein said isolated placental cells are amni on-derived adherent cells.
106. The method of claim 93, wherein said placental perfusate is combined with umbilical cord blood.
107. The method of claim 93, wherein said total nucleated cells from placental perfusate are combined with umbilical cord blood.
108. The method of claim 93, wherein said placental perfusate is combined with total nucleated cells from umbilical cord blood.
109. The method of claim 93, wherein said total nucleated cells from placental perfusate are combined with total nucleated cells from umbilical cord blood.
110. The method of claim 92, wherein said whole placenta is decellularized.
111. The method of claim 91 or claim 94, wherein said placental tissue is amniotic membrane.
112. The method of claim 111, wherein said amniotic membrane is decellularized.
113. The method of claim 91 or claim 94, wherein said placental tissue is chorionic membrane.
114. The method of claim 113, wherein said chorionic membrane is decellularized.
115. The method of claim 91 or claim 94, wherein said placental tissue is umbilical cord.
116. The method of claim 115, wherein said umbilical cord is decellularized.
117. The method of claim 91 or claim 94, wherein said placental tissue is umbilical cord vessels or placental vessels.
118. The method of claim 117, wherein said vessels are decellularized.
119. The method of claim 91 or claim 94, wherein said placental tissue is umbilical cord membrane.
120. The method of claim 119, wherein said umbilical cord membrane is decellularized.
121. The method of any of claims 91-120, wherein said banking of placental tissue is adjunct to banking of placental blood and/or umbilical cord blood.
122. The method of claim 91 or claim 93, wherein the method comprises banking of (i) at least one unit of placental blood or umbilical cord blood and (ii) at least one unit of perfusate or HPDSC, wherein said at least one unit of placental blood or umbilical cord blood, and said at least one unit of perfusate or HPDSC are from the same placenta.
123. The method of any of claims 91-120, wherein said placental tissue is banked without cryopreservation.
124. The method of any of claims 91-120, wherein said placental tissue is cryopreserved for banking.
125. The method of claim 124, wherein said placental tissue is cryopreserved by the placental tissue bank.
126. The method of any of claims 91-120, wherein said placental tissue is allogeneic to a customer of said placental tissue bank.
127. The method of any of claims 91-120, wherein said placental tissue is autologous to a customer of said placental tissue bank.
128. The method of any of claims 91-127, comprising assigning the placenta and/or placental tissue an identifier based on the source of the placenta.
129. The method of claim 128, comprising assigning the placenta a first identifier, and assigning each of said placental tissue a second tissue-specific identifier.
130. The method of claim 128, wherein said second tissue-specific identifier incorporates said first identifier.
131. The method of claim 129 or claim 130, wherein said placental tissue bank comprises a system for storing and retrieving said first and second identifiers.
132. The method of any of claims 91-94 wherein the method comprises receiving, prior to step (a) or step (b), an election from the customer as to which placental tissue from said placenta to bank.
133. The method of claim 132, wherein the placental tissue elected by the customer are assigned a third identifier to indicate election.
134. The method of claim 132 wherein placental tissue not elected by the customer are assigned a fourth identifier to indicate non-election.
PCT/US2016/022865 2015-03-18 2016-03-17 Methods of banking placental tissue and cells WO2016149492A1 (en)

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